Self-Perpetuating Structural States in Biology, Disease, and Genetics (2002) / Chapter Skim
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Identification of benzothiazoles as potential polyglutamine aggregation inhibitors of Huntington's disease by using an automated filter retardation assay
Identification of benzothiazoles as potential polyglutamine aggregation inhibitors of Huntington's disease by using an automated filter retardation assay
Pages 24-30
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From page 24... ...
We have developed an automated filter retardation assay for the rapid identification of chemical compounds that prevent HD exon 1 protein aggregation in vitro. Using this method, a total of 25 benzathiazole derivatives that inhibit huntingtin fibrillogenesis in a dose-dependent manner were discovered from a library of ~184,000 small molecules.
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From page 25... ...
HDQ51 (Flag-tagged HD exon 1 protein with 51 glutamines) expression was induced by thoroughly washing the cells with PBS and adding fresh medium lacking doxycycline.
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From page 26... ...
Recently, we have shown that formation of insoluble HD exon 1 protein aggregates in vitro is a nucleation-dependent process (32) and that addition of chemical compounds such as Congo red or thioflavine S to the aggregation reaction significantly delays the assembly of monomeric HD exon 1 protein into highmolecular-weight fibrillar structures (17~.
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From page 27... ...
(A) Effect of the indicated chemical compounds on HD exon 1 aggregation as monitored bythe filter retardation assay.
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From page 28... ...
Our data show that benzothiazole derivatives are potent inhibitors of HD exon 1 aggregation in vitro and in cell culture model systems of HD. However, the mechanism of action of these molecules on a molecular level is unclear.
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From page 29... ...
structurally related benzothiazole derivatives PGL-135 and PGL-137, identified by computer analysis, were nontoxic in the cell-based assay under the same conditions. We suggest that the compounds PGL-135 and PGL-137, similar to the most active compounds in the cell-free assay, directly interact with mutant HD exon 1 protein and thereby slow down the formation of insoluble protein aggregates in viva.
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From page 30... ...
The challenge is to find chemical compounds that are nontoxic, that have a reasonable brain permeability, and that prevent the formation of huntingtin aggregates in the patient's neurons. We have described a previously uncharacterized class of chemical compounds that prevent the formation of HD exon 1 aggregates in vitro and are biologically active in cell cultures.
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